DNA Technology
Recombinant DNA and
Genetic Engineering
Outline
1. Key Concepts
2. DNA Cloning
3. Genetic Engineering
4. Key Terms
5. Conclusions
Key Concepts
Genetic experiments have been
proceeding in nature for billions of years
Genetic changes are brought about by
Recombinant DNA technology
With technology, researchers can isolate,
cut, and splice together gene regions
from different species, and amplify the
number of copies
Key Concepts
Recombinant DNA technology depends on 3
activities
–
Cutting DNA into fragments
–
Insertion of fragments into cloning tools like plasmids
–
clone and identification of desired clone
Genetic engineering involves isolating,
modifying, and inserting genes back into the
same organism or into a different one
Social, ethical, legal, and ecological questions
are raised by the new technology
Glowing mice
Producing Restriction
Fragments
Restriction enzymes
–
Cut at specific nucleotide sequences
–
Some create “Sticky Ends”
–
DNA fragments cut with the same
restriction enzyme will base
-
pair to form
recombinant fragments
DNA ligase
–
Seals nicks where fragments base pair
Using a
restriction
enzyme and DNA
ligase to make
recombinant DNA
a. Restriction enzyme
cuts chromosomal or
cDNA
c. DNA or
cDNA
fragments
d. Plasmid
fragments
e. DNA fragments
and
modification
enzymes
are mixed
together
f. A collection of
recombinant
plasmids
g. Host cells able
to divide rapidly
take up
recombinant
plasmids
b. Same enzyme
cuts plasmid DNA
DNA Cloning and Genome (DNA) library
The original plasmid is called a
cloning vector
(taxi for delivering foreign DNA into a bacterium)
Human gene Cloning
Complementary DNA (cDNA)
Reverse
transcription
cDNA library
Polymerase Chain Reaction (PCR)
(any piece of DNA can be quickly amplified)
Heat DNA to unwind to single
strands
Add primers that base
-
pair with
ends
Cool mixture to promote base
-
pairing
DNA polymerase assembles
complementary sequences,
doubling number of identical
fragments
Repeat process over and over
Genetic engineering
The goal is to beneficially modifying traits
that the genes influence.
Bacteria
Genetically Engineered Bacteria
–
Insulin production
–
Growth hormone
–
Oil degraders
–
Pesticide control
Plant
Genetically
engineered cotton
plant having a gene
for herbicide
resistance.
Both had been
sprayed with a
weedkiller
Regenerating Plants from
Cultured Cells
Plant glows in
the dark owing
to a gene
transfer
Genetic Engineering of Animals
Insertion of gene into fertilized egg
Animals produce new products
Cloning
–
Dolly
Dolly and her first lamb
Applying the New Technology to
Humans
Human Genome Project
3.2 billion base pairs
–
99.9% of nucleotide sequence is same
for all humans
–
0.1% account for all genetic differences
DNA Sequencing
Genetic engineering
Recombination DNA
Biotechnology
Gene cloning
Restriction enzymes
cDNA
Cloning vector
Stick ends
DNA ligase
PCR
Genomic library
Polymerase chain reaction
Human Genome Project
In Conclusion
Nature conducts her own genetic engineering
experiments
Recombinant DNA technology allows the isolation,
cutting, and splicing of gene regions
Restriction enzymes and DNA ligase allow new DNA to
be inserted into plasmids
DNA libraries are collections of DNA fragments
Amplification can be done using bacteria or yeast or by
PCR
In Conclusion
Genetic engineering involves the isolation,
modification, and insertion of genes
Recombinant DNA technology and genetic
engineering have potential research and
applications in medicine, agriculture, and
industry
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